Methods of preparing and using novel stem cell compositions and kits comprising the same
The present invention provides novel stem cell compositions having significant therapeutic and practical advantages, as well as methods of preparing and using such compositions for the treatment and prevention of injury and disease in patients. The invention may be applied to stem cell populations isolated from a wide variety of animals, including humans, and tissues. In particular applications, the invention is used to prepare a stem cell composition from a collagen-based tissue, such as adipose tissue, isolated from a patient, and the stem cell composition is subsequently provided to a site of actual or potential injury in the patient. The invention further includes related kits comprising the stem cell compositions, which are remarkably stable and retain viability and efficacy during storage and shipment.
1. A method of treating inflammation at a site of a musculoskeletal injury or disease in a mammal, comprising:
(a) preparing a cell population comprising adipose tissue-derived stem cells for introducing into a mammal by a method comprising:
(i) processing adipose tissue obtained from the mammal to release cells therein, wherein the adipose tissue comprises stem cells, wherein said processing comprises one or more procedures selected from the group consisting of:
physical cutting or mincing of the adipose tissue;
treating the adipose tissue with an enzyme that facilitates the release of cells;
exposing the adipose tissue to ultrasonic energy; and
treating the adipose tissue with perfluorocarbons,
and wherein said processing releases cells and a fat layer; and
(ii) separating the cells released in (i) from the fat layer, wherein said separating comprises one or more procedures selected from the group consisting of:
use of a density gradient;
centrifugation; and
filtration;
thereby preparing the cell population comprising adipose tissue-derived stem cells; and
(b) providing the cell population of (a)(ii) directly to the site of the musculoskeletal injury or disease in the mammal to treat the inflammation at the site of the musculoskeletal injury or disease,
wherein said method does not include isolating stem cells separated in (a)(ii) from other cells separated in (a)(ii).
2. The method of claim 1 , wherein said cell population further comprises one or more cells selected from the group consisting of: red blood cells, white blood cells, fibroblasts, neutrophils, monocyte/macrophages, and basophils.
3. The method of claim 1 , wherein said cell population further comprises one or more tissue components selected from the group consisting of: extracellular matrix polypeptides, proteoglycans, cytokines, and growth factors.
4. The method of claim 1 wherein the mammal has a tail, and said adipose tissue is obtained from the tail head region of the mammal.
5. The method of claim 1 , further comprising:
(iii) suspending the separated cells in a physiologically compatible buffer.
6. The method of claim 1 , further comprising:
(iii) placing the separated cells in a container.
7. The method of claim 6 , wherein the container is a syringe.
8. The method of claim 6 , further comprising shipping the container comprising the separated cells to a physician or veterinarian.
9. The method of claim 1 , wherein the musculoskeletal injury or disease is a sprain, strain, dislocation, bruising, tear, microtear, contusion, bursitis, tendonitis, articular cartilage injury or fracture.
10. The method of claim 1 , wherein the musculoskeletal injury or disease is in a tendon, ligament, cartilage, bone, or hoof laminae.
11. The method of claim 1 , wherein the mammal is a human, a dog, a cat, a horse or a camel.
12. The method of claim 11 , wherein the mammal is a horse.
13. The method of claim 9 , wherein the musculoskeletal injury or disease is tendonitis.
14. The method of claim 1 , wherein the treatment of the inflammation comprises a reduction in inflammatory cell infiltrates at the site of the musculoskeletal disease or injury.
15. The method of claim 1 , wherein: the mammal is a human, a dog, a cat, a horse or a camel; the musculoskeletal injury or disease is tendonitis; and the treatment of the inflammation comprises a reduction in inflammatory cell infiltrates at the site of injury.
16. The method of claim 1 , wherein the inflammation is at the site of the musculoskeletal injury.
17. The method of claim 16 , wherein the musculoskeletal injury is a sprain, strain, dislocation, bruising, tear, microtear, contusion, bursitis, tendonitis, articular fracture in cartilage injury or fracture in a tendon, ligament, cartilage, bone, or hoof laminae.
18. The method of claim 16 , wherein the site of the musculoskeletal injury is a tendon, ligament, cartilage, bone, or hoof laminae.
19. The method of claim 1 , wherein the inflammation results from the musculoskeletal injury or disease.